Human Dystroglycan Antibody AF6868: R&D Systems

Human Dystroglycan Antibody

(1 citations)   
  • Species Reactivity
    Human
  • Specificity
    Detects human Dystroglycan in direct ELISAs and Western blots.
  • Source
    Polyclonal Sheep IgG
  • Purification
    Antigen Affinity-purified
  • Immunogen
    Mouse myeloma cell line NS0-derived recombinant human Dystroglycan
    Gln28-Val749
    Accession # Q14118
  • Formulation
    Lyophilized from a 0.2 μm filtered solution in PBS with Trehalose. *Small pack size (SP) is supplied as a 0.2 µm filtered solution in PBS.
  • Label
    Unconjugated
Applications
  •  
    Recommended
    Concentration
    Sample
  • Western Blot
    1 µg/mL
    See below
  • Immunohistochemistry
    5-15 µg/mL
    See below
Please Note: Optimal dilutions should be determined by each laboratory for each application. General Protocols are available in the Technical Information section on our website.
Data Examples
Detection of Human Dystroglycan by Western Blot. Western blot shows lysates of MCF‑7 human breast cancer cell line, SH‑SY5Y human neuroblastoma cell line, human muscle tissue, and human placenta tissue. PVDF membrane was probed with 1 µg/mL of Sheep Anti-Human Dystroglycan Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6868) followed by HRP-conjugated Anti-Sheep IgG Secondary Antibody (Catalog # HAF016). Specific bands were detected for alpha ‑Dystroglycan at approximately 100-160 kDa (as indicated) and beta ‑Dystroglycan at approximately 42-44 kDa (as indicated). This experiment was conducted under reducing conditions and using Immunoblot Buffer Group 1.
Immunohistochemistry
Dystroglycan in Human Skeletal Muscle. Dystroglycan was detected in immersion fixed paraffin-embedded sections of human skeletal muscle using Sheep Anti-Human Dystroglycan Antigen Affinity-purified Polyclonal Antibody (Catalog # AF6868) at 3 µg/mL overnight at 4 °C. Before incubation with the primary antibody, tissue was subjected to heat-induced epitope retrieval using Antigen Retrieval Reagent-Basic (Catalog # CTS013). Tissue was stained using the Anti-Sheep HRP-DAB Cell & Tissue Staining Kit (brown; Catalog # CTS019) and counterstained with hematoxylin (blue). Specific staining was localized to basement membrane. View our protocol for Chromogenic IHC Staining of Paraffin-embedded Tissue Sections.
Preparation and Storage
  • Reconstitution
    Sterile PBS to a final concentration of 0.2 mg/mL.
  • Shipping
    The product is shipped at ambient temperature. Upon receipt, store it immediately at the temperature recommended below. *Small pack size (SP) is shipped with polar packs. Upon receipt, store it immediately at -20 to -70 °C
  • Stability & Storage
    Use a manual defrost freezer and avoid repeated freeze-thaw cycles.
    • 12 months from date of receipt, -20 to -70 °C as supplied.
    • 1 month, 2 to 8 °C under sterile conditions after reconstitution.
    • 6 months, -20 to -70 °C under sterile conditions after reconstitution.
Background: Dystroglycan

Dystroglycan, also DAG-1 (Dystrophin-associated glycoprotein 1) is a 180-200 kDa heterodimeric adhesion molecule that links the cell cytoskeleton to the extracellular matrix. It is found on skeletal muscle, cardiac muscle, fibroblasts, smooth muscle and keratinocytes. DAG-1 binds multiple matrix molecules, including laminin-1 and -2, agrin, and perlecan. Intracellularly, the cytoplasmic tail of DAG-1 contributes to a large 400 kDa complex that interacts with the cytoskeleton. The human DAG-1 preprocursor is a type I transmembrane protein 895 amino acids (aa) in length. It contains a 27 aa signal sequence plus an 868 aa proform that undergoes autocatalysis to generate a 626 aa alpha -chain (aa 28-653), and a 242 aa beta -chain. Mature DAG-1 is a heterodimer composed of noncovalently linked alpha ‑ and beta ‑chains. The alpha -chain possesses one potential Ig-like domain (aa 64-162), a mucin-like region (aa 316-485), and a peptidase S72 domain (aa 500-733). It is O‑glycosylated and runs from 100-160 kDa in SDS-PAGE. The beta -chain is N-glycosylated and runs at 42-44 kDa in SDS-Page. It possesses a short 95 aa extracellular region (aa 654-749) plus a 120 aa cytoplasmic domain (aa 776-895). Membrane cleavage of the beta -chain causes dissociation of the heterodimer and generates a 30 kDa truncated form. Over aa 28-749, human DAG-1 shares 93% aa identity with mouse DAG-1.

  • Long Name:
    Dystrophin-associated Glycoprotein 1
  • Entrez Gene IDs:
    1605 (Human); 13138 (Mouse); 114489 (Rat)
  • Alternate Names:
    A3a; AGRNR; DAG1; Dag-1; DAG156DAG; dystroglycan 1 (dystrophin-associated glycoprotein 1); Dystroglycan; Dystrophin-associated glycoprotein 1
Related Research Areas
Citations:

R&D Systems personnel manually curate a database that contains references using R&D Systems products. The data collected includes not only links to publications in PubMed, but also provides information about sample types, species, and experimental conditions.

1 Citations: Showing 1 - 1

  1. Structural basis of laminin binding to the LARGE glycans on dystroglycan
    Nat Chem Biol, 2016;0(0):.
    Species: Rabbit
    Sample Type: Protein
    Application: WB
Cell and Tissue Staining Kits
Description Application Cat# Citations Images  

Anti-Sheep HRP-DAB Cell & Tissue Staining Kit

CTS019 1
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Immunohistochemistry Reagents
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Antigen Retrieval Reagent-Universal

CTS015 10
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Antigen Retrieval Reagent-Basic

CTS013 4
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Mounting Medium

CTS011 4
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Antigen Retrieval Reagent Sampler (50 mL each)

CTS016
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Antigen Retrieval Reagent-Acidic

CTS014
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NorthernLights Guard Mounting Media

NL996
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VisUCyte Antigen Retrieval Reagent-Acidic

VCTS022
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VisUCyte Antigen Retrieval Reagent-Universal

VCTS023
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VisUCyte Antigen Retrieval Reagent-Basic

VCTS021
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Isotype Controls
Description Application Cat# Citations Images  

Normal Sheep IgG Control

Ctrl 5-001-A 9
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Secondary Antibodies
Description Application Cat# Citations Images  

Sheep IgG HRP-conjugated Antibody

WB, Simple Western HAF016 14  
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Donkey Anti-Sheep IgG NorthernLights™ NL557-conjugated Antibody

Flow, IHC, ICC NL010 3
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Donkey Anti-Sheep IgG NorthernLights™ NL637-conjugated Antibody

Flow, IHC, ICC NL011 5
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Donkey Anti-Sheep IgG NorthernLights™ NL493-conjugated Antibody

Flow, IHC, ICC NL012 2
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Sheep IgG (H+L) PE-conjugated Antibody

Flow F0126  
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Donkey Anti-Sheep IgG Biotinylated Antibody

WB BAF016 2
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Sheep IgG (H+L) APC-conjugated Antibody

Flow F0127  
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Sheep IgG VisUCyte HRP Polymer Antibody

IHC VC006  
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Donkey Anti-Sheep IgG (H+L) Fluorescein-conjugated Antibody

Flow F0125 1
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Donkey Anti-Sheep IgG (H+L) PerCP-conjugated Antibody

Flow F0128
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